biology essay

Biochemical And Haematological Parameters Biology Essay

Published: 23, March 2015

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The biochemical and haematological effects of the seed powder of Mucuna pruriens in male rats were evaluated to establish some biological properties of this potential biopesticide currently undergoing investigation. The result showed that Mucuna pruriens seed extract produced a significant (P&lt;0.05) increase in white blood cell (WBC) count, as well as in bilirubin concentrations, alkaline phosphatase (ALP), protein and creatinine levels measured. Uric acid, alanine aminotransferase (ALT) and aspartate aminotransferase (AST) are significantly reduced (P&lt;0.05) in comparison with the experimental control. RBC, Hb, PCV, BUN, Protein and Albumin levels gave no significant difference between treated and control group. The results revealed metabolic imbalance in the rats which suggests a poor renoprotective effect compared to previous works.

INTRODUCTION

Enzymes are proteinous biomolecules that catalyze metabolic reactions (Grisham and Reginald, 1999); they are responsible for anabolic and catabolic activities in biosystems. Significantly elevated levels, for example, of Alanine aminotransferase (ALT) in the plasma signifies extensive hepatic activity and possible damage. Similarly, haematological parameters are used as laboratory indices to evaluate events in biosystems; the WBC count indicates the immune status and haematologic integrity of the body defense mechanism.

Mucuna pruriens seed is claimed to be used as a preservative for seed storage in some parts of Plateau and Benue states in Nigeria. It is an annual, climbing shrub with long vines that can reach over 15 m. It bears white, lavender, or purple flowers and pods that are covered in loose orange hairs which cause a severe itch if they come in contact with skin. They are shiny black or brown drift seeds, found in tropical Africa, India and the Caribbean. The Mucuna pruriens seeds are demonstrated to contain high concentrations of levodopa, a direct precursor of the neurotransmitter dopamine. This explains why this plant has long been used in traditional Ayurvedic Indian medicine for diseases including parkinson's disease (Manyam et al., 2004). It has also been documented that the plant is useful in the treatment of a lot of ailments including the prevention of coagulation effects of snake bites (Aguiyi et al., 1996). This study is aimed at evaluating the effects of Mucuna pruriens seed extract on haematological and biochemical parameters using adult male rats.

MATERIALS AND METHODS

Animals

Adult albino male rats of the Wistar strain, weighing between 130-150 g were obtained from the Animal House facility of the University of Jos, Nigeria. These animals were housed in a well ventilated and spacious room (room temperature 27+3oC). All animals were fed with standard diet prepared at the Animal House facility, and were given water ad libitum.

Collection and Identification of Plant Seeds

Seeds of Mucuna pruriens were cultivated and obtained from Benue state, Nigeria and were identified by Professor C.O. Akueshi of the Department of Botany, University of Jos, Nigeria.

Extraction Procedure

The crude extract was prepared by adding 350 ml of deionized water to 35 g of Mucuna pruriens extract and exhaustive extraction for 72 hours at a temperature range of 65-70oC, to obtain hot water extracts (HWE) of Mucuna pruriens. Percentage yield of the extract obtained was derived and pH of extract was measured using a pH meter.

Toxicology of Plant Seeds

Adult male Wistar strain rats weighing between 130-150 g were used for acute toxicity studies and the sub-chronic studies. Adopting the method developed by (Lorke, 1983), acute toxicity testing was done for Mucuna pruriens using the male rats. The LD50 was obtained for Mucuna pruriens in the rats used. In the case of sub-chronic studies, two dose points of 500 mg/kg and 1000 mg/kg for Mucuna pruriens seed powder were employed. Thirty-six (36) rats of 130-150 g weight distribution were put into labeled cages A to F and separated into six (6) rats per group, and rats in first three cages were each administered 500 mg/kg of Mucuna pruriens. The rats in the remaining three cages were each administered 1000 mg/kg of Mucuna pruriens. Administration of the extract was daily and lasted for six weeks. Biochemical and haematological parameters were then evaluated at the end of the six weeks, using the method of Tienz (1970).

STATISTICAL ANALYSIS

All treatments were performed in triplicate and each data point in the results is the mean of two or three replicate tests. All experiments were repeated at least once. The statistical significance of a treatment effect was evaluated by student's t-test and the values were ex-pressed as mean ± SEM (standard error of mean).

RESULTS

Percentage yield of Mucuna pruriens extract was 21.57% and pH of extract measured gave 8.76.

The LD50 was obtained for Mucuna pruriens in male rats as 1300 mg/kg body weight.

Table 1 shows the measured effect of extract of Mucuna pruriens on some biochemical parameters in male rats.

Table 1: Effect of Extract of Mucuna pruriens on some Biochemical Parameters in Male Rats

Treatment

Protein

(g/l)

Albumin

(g/l)

ALT

(UI)

AST

(UI)

ALP

(UI)

Control (untreated seeds)

70.46±0.14

44.50±0.17

40.00±0.58

131.33±0.88

445.23±0.07

Mucuna

(500 mg/kg)

80.43±0.20*

40.43±0.12#

19.00±0.58#

78.66±0.88#

520.60±0.06*

Mucuna

(1000 mg/kg)

98.66±0.33*

54.80±0.05*

18.33±0.62#

95.00±1.15#

581.46±0.18*

Significant difference, at *P&lt;0.05 and #P&gt;0.05 for treatment versus control

Control - Untreated weevil-infested Vigna unguiculata seeds

ALT - Alanine aminotransferase

AST - Aspartate aminotransferase

ALP - Alkaline Phosphatase

Table 2 shows the measured effect of extract of Mucuna pruriens on some biochemical parameters in male rats.

Table 2: Effect of Extract of Mucuna pruriens on some Biochemical Parameters in Male Rats

Treatment

T.Bilirubin (mMol/L)

D.Bilirubin

(mMol/L)

Glucose (mMol/L)

Creatinine (mMol/L)

Control

(untreated seeds)

1.46±0.03

0.63±0.03

7.63±0.17

145.53±0.12

Mucuna

(500 mg/kg)

3.80±0.08*

2.56±0.03*

6.50±0.15#

144.60±0.17#

Mucuna

(1000mg/kg)

0.83±0.06#

0.50±0.03#

4.53±0.15#

165.53±0.20*

Significant difference, at *P&lt;0.05 and #P&gt;0.05 for treatment versus control

T. Bilirubin - Total Bilirubin

D. Bilirubin - Direct Bilirubin

Table 3 shows the measured effect of extract of Mucuna pruriens on some hematological parameters in male rats.

Table 3: Effect of Extract of Mucuna pruriens on some Haematological Parameters in Male Rats

Treatment

PCV

(%)

HB

(d/l)

WBC

(cell/mm2)

LYMPH

(%)

Control

(untreated seeds)

36.67±0.8

13.68±0.9

6233.33±16.6

83.33±0.8

Mucuna

(500 mg/kg)

40.33±0.3#

15.46±0.2#

26250.00±28.8*

83.00±0.6#

Mucuna

(1000mg/kg)

46.00±0.5#

16.92±0.3#

6516.67±16.1*

86.00±0.6#

Significant difference, at *P&lt;0.05 and #P&gt;0.05 for treatment versus control, using Student's t-test, (n=6 per group). Values are expressed as mean ± S.E.M

PCV - Packed Cell Volume

HB - Haemoglobin concentration

WBC - White Blood Cells

LYMPH - Lymphocytes

Table 4 shows the measured effect of extract of Mucuna pruriens on some hematological parameters in male rats.

Table 4: Effect of Extract of Mucuna pruriens on some Haematological Parameters in Male Rats

Treatment

NEUTR

(%)

EOSINO

(%)

MON

(%)

BFP

Control

(untreated seeds)

16.00±0.58

0

0.33±0.3

Normal

Mucuna

(500 mg/kg)

16.00±0.58#

0.33±0.3#

0

Normal

Mucuna

(1000mg/kg)

13.00±0.58#

0.33±0.3#

0

Normal

Significant difference, at *P&lt;0.05 and #P&gt;0.05 for treatment versus control

NEUTR - Neutrophils

EOSINO - Eosinophils

MON - Monophils

BFP - Blood Film Picture

DISCUSSION

The measured pH of 8.76 for Mucuna pruriens revealed its tilt toward alkalinity which generally would not affect the action of the extract in vivo. There was significant difference (P&lt;0.05) between treated and control groups, for both haematologic parameters assessed and biochemical enzymes assayed from serum samples collected from the male rats sacrificed after six weeks of daily administration of extract. However, there was a significant difference in WBC total count of rats administered with Mucuna pruriens as seen in Table 3, compared with the control. A higher average WBC total count of 26,000 cells/mm2 was measured. This significant increase (P&lt;0.05) in WBC total count was probably triggered off by the metabolic assault from alkaloidal or phenolic content in Mucuna pruriens (Rajaram and Janardhanan, 1991). L-dihydroxyphenylacetic acid (L-dopa) is another key constituent in Mucuna pruriens that may likely be responsible for such observed findings in WBC total counts (Rajaram and Janardhanan, 1991).

The membrane bound target enzymes Aspartate aminotransferase (AST) and Alanine aminotransferase (ALT) significantly decreased in liver tissues when measured after six weeks of treatment. A high degree positive correlation was observed with regard to these enzymes between serums versus kidney, whereas in case of serum versus liver tissues high degree negative correlation was recorded. These enzyme profiles elucidate that they decreased significantly in liver indicating necrosis of these tissues (Rahman and Siddiqui, 2003), which is suggestive that an increase synthesis of these enzymes, may be an adaptive mechanism due to the stress of the toxicant. These biomarker enzymes can be detected rapidly and hence may be used for the prediction and diagnosis of metabolic insults. Average values of ALT and AST measured for Mucuna pruriens seed powder administered in rats produced a decrease compared to the control. Mucuna pruriens is a known antioxidant (Aguiyi et al., 1996; Tripathi and Upadhyay, 2001), therefore, the suppression of liver enzymes to significant values, could be explained by the enhanced suppressive effect displayed by Mucuna pruriens extract in preventing enzymatic over-sensitization to the metabolism of various substances foreign to the normal system in the rats used for the study.

CONCLUSION

Haematological and biochemical investigations carried out showed no significant difference (P&gt;0.05) in the parameters measured for RBC, Hb, PCV, BUN, Protein and Albumin levels. However, Mucuna pruriens caused an increase in total WBC count that was significant (P&lt;0.05), as well as in ALP, protein, creatinine levels and bilirubin concentrations measured. Uric acid, Alanine aminotransferase and Aspartate aminotransferase levels were reduced in comparison with their controls respectively.

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